U.S. patent number 10,357,523 [Application Number 14/400,337] was granted by the patent office on 2019-07-23 for animal feed comprising grain and agaricus blazei extract and use of the feed material.
This patent grant is currently assigned to SSIPFEED B.V.. The grantee listed for this patent is Pierre Michel Grammare, SSIPfeed B.V., Wilhelmus Hubertus Henricus Antonius Van Den Elshout. Invention is credited to Pierre Michel Grammare, Wilhelmus Hubertus Henricus Antonius Van Den Elshout.
United States Patent |
10,357,523 |
Van Den Elshout , et
al. |
July 23, 2019 |
Animal feed comprising grain and agaricus blazei extract and use of
the feed material
Abstract
The invention relates to the use of feed material comprising
grain and Agaricus Blazei, comprising the Agaricus species in an
amount between 10 to 50% by weight on dry matter, and the feed
material having a moisture content of less than 10% (relative to
dry matter), for administration to chicken for improvement of egg
laying, preferably for improving shell quality and/or for extending
the egg laying period. The invention further relates to a process
for making the feed material, and feed for laying hens comprising
the feed material.
Inventors: |
Van Den Elshout; Wilhelmus Hubertus
Henricus Antonius (Venlo, NL), Grammare; Pierre
Michel (Lakanal, FR) |
Applicant: |
Name |
City |
State |
Country |
Type |
Van Den Elshout; Wilhelmus Hubertus Henricus Antonius
Grammare; Pierre Michel
SSIPfeed B.V. |
Venlo
Lakanal
Venlo |
N/A
N/A
N/A |
NL
FR
NL |
|
|
Assignee: |
SSIPFEED B.V. (Venlo,
NL)
|
Family
ID: |
48463970 |
Appl.
No.: |
14/400,337 |
Filed: |
May 14, 2013 |
PCT
Filed: |
May 14, 2013 |
PCT No.: |
PCT/EP2013/059903 |
371(c)(1),(2),(4) Date: |
November 11, 2014 |
PCT
Pub. No.: |
WO2013/171194 |
PCT
Pub. Date: |
November 21, 2013 |
Prior Publication Data
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|
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Document
Identifier |
Publication Date |
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US 20150140098 A1 |
May 21, 2015 |
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Foreign Application Priority Data
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|
|
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May 14, 2012 [NL] |
|
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2008812 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
31/07 (20130101); A61K 36/07 (20130101); A23K
10/12 (20160501); A23K 20/163 (20160501); A61K
31/575 (20130101); A61K 31/201 (20130101); A23K
20/174 (20160501); A61K 31/59 (20130101); A23K
10/30 (20160501); A23K 50/75 (20160501) |
Current International
Class: |
A23K
10/12 (20160101); A23K 10/30 (20160101); A61K
31/07 (20060101); A61K 31/59 (20060101); A61K
31/201 (20060101); A23K 50/75 (20160101); A61K
31/575 (20060101); A61K 36/07 (20060101); A23K
20/163 (20160101); A23K 20/174 (20160101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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|
|
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|
|
101371683 |
|
Feb 2009 |
|
CN |
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1714674 |
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Oct 2006 |
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EP |
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2004/321033 |
|
Nov 2004 |
|
JP |
|
20050013900 |
|
Feb 2005 |
|
KR |
|
2097979 |
|
Dec 1997 |
|
RU |
|
2542523 |
|
Feb 2015 |
|
RU |
|
Other References
Sanchez, C "Reactive oxygen species and antioxidant properties from
mushrooms" Synthetic and Systems Biotechnology 2017, 2, 13-22.
DOI:10.1016/j.synbio.2016.12.001. cited by examiner .
Jonathan et al "Effect of physical and chemical factors on mycelial
growth of ten wild Nigerian mushrooms during cellulase and amylase
production" NPAIJ, 2011, 7(4),p. 211-216. cited by examiner .
Block, SS et al "Experiments with Submerged Culture" Agricultural
and Food Chemistry, Sep. 30, 1953, p. 890-893. cited by examiner
.
Amerah AM et al "Feed particle size: Implications on the digestion
and performance of poultry" World's Poultry Science Journal, 2007,
63,p. 439-455. DOI: 10.1017/S0043933907001560. cited by examiner
.
Unicorn Imp & Mfg Corp ("Cultivation of Agaricus blazei"
retrieved online
<URL:https://unicornbags.com/cultivation/agaricus-blazei>,
accessed Sep. 2017 (archived May 2, 2012), 14 pages. cited by
examiner .
Ikeya M, et al "The effect of feeding ground Agaricus blazei Murill
on egg production in hen", Bull. of Shizuoka Swine and Poultry
Exper. Station, The Ag., Forestry and Fisheries Res. Info. Technol.
Ctr., AFFRIT, Japan, No. 14, Jan 1, 2003, pp. 29-32,, ISSN:
0914-6520 (Eng. Translation, 12pp.) (Year: 2003). cited by examiner
.
Dutch Search Report for priority document NL2008812 dated Feb. 5,
2013. cited by applicant .
International Search Report for PCT/EP2013/059903 dated Jun. 28,
2013. cited by applicant .
International Preliminary Report on Patentability for
PCT/EP2013/059903 dated Nov. 18, 2014. cited by applicant .
Ikeya M et al: "The effect of feeding ground Agaricus blazei Murill
on egg production in hen", Bulletin of Shizuoka Swine and Poultry
Experiment Station, The Agriculture, Forestry and Fisheries
Research Information Technology Center, Affrit, Japan, nr. 14, Jan.
1, 2003, pp. 29-32, XP008159889, ISSN: 0914-6520. cited by
applicant .
Jonathan, Segun G. et al: "Evaluation of Ten Wild Nigerian
Mushrooms for Amylase and Cellulase Activities", Mycobiology 39(2):
103-108 (2011). cited by applicant .
Francois, Jean M: "A simple method for quantitative determination
of polysaccharides in fungal cell walls", Nature protocals (2007)
1:2995-3000. cited by applicant.
|
Primary Examiner: Kosar; Aaron J
Attorney, Agent or Firm: Amirsehhi; Ramin Owen; David P.
Tsai; Philip
Claims
The invention claimed is:
1. A process for producing a feed material, consisting of: a)
fermenting mycelium of Agaricus blazei fungus on an amount of
grain, wherein the moisture content by weight of the grain is
between 10 wt % and 90 wt %, until the fermented grain comprises by
dry weight thereof 15 wt % to 50 wt % mycelium content, b) drying
the fermented grain at a temperature between 25.degree. C. and
90.degree. C. and to reduce the moisture content thereof of less
than 10 wt %, and c) milling the so obtained fermented and dried
material to a particle size with a d.sub.50 between 0.01 mm and 10
mm thereby producing said feed material.
2. The process according to claim 1, wherein the grain is rye or
oat.
3. The process according to claim 1, wherein the dried material has
a particle size with a d.sub.50 between 0.01 mm and 3 mm.
4. The process according to claim 1, wherein the drying temperature
is below 50.degree. C.
5. The process according to claim 1, wherein the feed material has
a moisture content between 2 wt % and 8 wt %.
6. The process according to claim 1, wherein the feed material has
a particle size with a d.sub.50 between 0.01 mm and 3 mm.
7. The process according to claim 1, wherein the feed material
comprises ergosterol in an amount of 100 mg and 600 mg per kg feed
material.
8. The process according to claim 1, wherein the feed material
comprises active cellulases having an activity between 0.1 to 0.8
unit/ml in an extract which is obtained if 10 g feed material is
extracted with 20 ml water.
9. The process according to claim 1, wherein the feed material
comprises an amount of 1,3- and 1,6-.beta.-glucans of between 3 g
and 100 g, per kg of the feed material.
10. The process according to claim 1, wherein the feed, per kg
thereof, comprises: an amount of ergosterol of 100 mg to 600 mg,
and an amount of 1,3- and 1,6-.beta.-glucans of between 3 g and 100
g per kg of the feed material.
11. The process according to claim 1, wherein the grain is rye or
oat, wherein the dried material has a particle size with a d.sub.50
between 0.01 mm and 3 mm, and wherein the feed material has a
moisture content between 2 wt % and 8 wt %.
Description
CROSS REFERENCE TO RELATED APPLICATION
This application is a 35 U.S.C. 371 National Phase Entry
Application from PCT/EP2013/059903, filed May 14, 2013, which
claims the benefit of Dutch Patent Application No. 2008812 filed on
May 14, 2012, the disclosure of which is incorporated herein in its
entirety by reference.
This invention relates to feed material for chicken and other
fodder, and more in particular to the use of this feed material for
improving long term egg shell quality, and increasing the egg
laying period of laying hens.
Chicken and eggs are a widely used as food for human consumption.
Large numbers of chicken are grown for egg production. Generally,
female chicken (laying hens) are grown for 17 weeks, after which
period they can be used for egg production. The growth and
maintenance of the laying hens is highly optimized, and currently,
the hens are producing eggs for about 60 weeks. Initially, the
laying percentage reaches about 95% (>93%) after about 10 weeks,
for a period of about 10 weeks, but thereafter the laying
percentage decreases. Also, the quality of the egg shells
deteriorates on average.
It is an object of the invention to provide a feed, or feed
material, that allows an improvement in egg shell quality and/or
egg laying period for the laying hens.
The object of the invention is achieved with providing the chickens
with a feed or feed material comprising Agaricus blazei, wherein
the Agaricus species is fermented on grain, the fermented grain is
dried, and optionally pulverized, and used as feed material. Hence,
the present invention relates to a feed material comprising grain
and Agaricus blazei, comprising the Agaricus species in an amount
between 10 to 50% by weight on dry matter, and the feed material
having a moisture content of less than 10% (relative to dry
matter), for administration to chicken for improvement egg laying,
preferably for improvement of egg shell quality and/or for
extending the egg laying period.
The mushroom species according to the invention is an Agaricus
blazei, preferably Agaricus blazei murill (also called A. blazei
Brasiliensis, or Agaricus subrufescens, or Agaricus rufotegulis).
At present, it is thought that Agaricus subrufescens is the correct
name; however, in this application, the more common Agaricus blazei
murill will be used. Herein below, the abbreviation ABM for
Agaricus blazei murill is used interchangeably. The preferred
Agaricus species is Agaricus blazei murill.
The use of extracts of Agaricus blazei for administration to laying
hens, for example an extract in water, as a solution or absorbed on
zeolites, is described in JP2004/321033. Some improvement is seen
in strengthening the immune system according to this reference.
However, JP2004/321033 does not suggest the improvements of the
present invention. Further, Ikeya et al. (Bulletin of the Shizuoka
swine and poultry experiment station, the agriculture, forestry and
fishery research information technology center (AFFRIT), Japan, no
14, 1 Jan. 2003 pp 29-32 (XP008159889, ISSN 0914-6520)) describes
the use of ground Agaricus blazei murill for laying hens. However,
the improvements found in the present invention are neither
described, nor suggested therein.
Fermentation of Agaricus blazei species on grain is known as such,
also for feed applications in general, like for example described
in CN101371683. However, the improvements found in the present
invention are neither described, nor suggested therein.
US2008/187574 describes the fermentation of multiple microorganisms
on grain. However, the improvements found in the present invention
are neither described, nor suggested therein.
The mushroom species according to the invention, which gives very
good results, is an Agaricus blazei, preferably Agaricus blazei
murill (also called A. blazei Brasiliensis). Herein below, the
abbreviation ABM for Agaricus blazei murill will be used
interchangeably.
The Agaricus blazei fungus is grown as mycelium on a grain source,
and harvested as such. Hence, it is not necessary to execute
mycelium growth till fruit bodies emerge, nor is it necessary to
separate the growth medium from the mycelium. Thereby, the present
invention allows for an economically effective way of providing
useful feed for chicken resulting in increased egg production. The
improvement is seen during the normal egg laying period, but is
also clear from improved egg shell, and increased laying
period.
As the grain source, several commonly known grain types are useful,
such as for example corn, wheat bran, oat, sorghum, barley, whole
wheat, rye, soy beans, maize and the like. Mixtures can be used as
well. Further, addition of carbon source, ammonia source or the
like can be useful to increase growth of the mycelium. Calcium
compounds like chalk may be added. Rye or oat are particularly
preferred because ABM grows particularly well on these
substrates.
In order to obtain a suitable fermented ABM, it is preferred that
the grain source comprises a sufficiently high moisture content. A
suitable moisture content is between about 10-80% moisture
(measured as weight of the moist product minus a dried product,
divided by the moist product weight). A preferred moisture content
is between about 30 and 70%, like for example about 50%.
The grain source is sterilized before inoculation with the ABM
mycelium. Inoculation, and the preparation of the inoculate follows
standard techniques, like for example described in U.S. Pat. No.
4,204,364.
The fermentation generally takes place in containers of 20 to 50
liter size, like in bags or trays, like for example 25 or 30 liter
bags. Fermentation preferably takes place in a conditioned
environment. Generally, the time for fermentation will be between
15 and 75 days, like between 15 and 55 days, preferably between 20
and 45 days. Too short will cause relatively low mycelium content;
a too long period is economically less interesting. The temperature
during fermentation preferably is between 20 and 35.degree. C., and
most preferred between 28 and 30.degree. C. The humidity preferably
is between 40% and 90% RH, such as for example 50 or 60% RH.
After a suitable period of fermentation, the amount of mycelium is
between about 10 and 50% (on dry weight of the mixture of grain and
mycelium), preferably between 20 and 30%. The amount of mycelium
can be measured indirectly, for example based on the ergosterol
content, as will be further elucidated in the examples. Any
suitable method for determining the mycelium content can be used.
For example, it is also possible to measure the mycelium content
based on the amount of chitin.
After the fermentation, the fermented grain is dried. The moisture
content of the dried fermented grain preferably is about 10% or
less, preferably about 8% or less and more preferably about 6% or
less. A lower moisture content aids in achieving a good storage
stability. Generally, the moisture content will be about 2% or
more, and preferably about 3% or more. As such, a feed with very
low moisture content has no disadvantages from technical
perspective, but it is more costly to produce.
Any drying technique may be suitable. A suitable apparatus includes
a belt dryer, bulb dryer, tumble dryer or fluid bed dryer.
Preferably, the apparatus is such that it is able to perform the
drying at reduced pressure. In general, it is preferred that the
temperature remains below 100.degree. C. Preferably, the fermented
grain is dried by air drying at between 5 and 100.degree. C.,
preferably between 25 and 90.degree. C., more preferably below
63.degree. C., and even more preferably below 50.degree. C. Drying
at a temperature below about 50.degree. C. has the advantage that
extra-cellular enzymes, like cellulases keep their activity. In
case the enzymes that are of interest have been modified to be
resistant to higher temperatures, a higher temperature for drying
is preferred because drying speed is increased. If a temperature is
chosen below 63.degree. C., 45.degree. C., or in particular at
about 35.degree. C. or lower, it is preferred to apply reduced
pressure. The low temperature applied during the drying step has
the further advantage that heat sensitive compounds produced during
fermentation remain in an active form in the feed.
The dried fermented grain is storage stable for several months up
to at least one year (e.g. for two years), with little reduction of
the nutritive value of the feed material.
The fermented dried grain (feed material) can be used as such, as
or in chicken feed.
For improving mixing characteristics with common feedstock, and to
allow reproducible administration to the laying hens, it is
preferred to crush or mill the fermented dried grain to a particle
size (d.sub.50) between 0.01 and 10 mm, preferably lower than 5 mm,
and even more preferably between 0.1 and 3 mm. The particle size
range preferably is between 0.1 and 2.0 mm for about 90% or more of
the feed material, preferably of about 95% or more of the feed
material, and more preferably of about 98% or more of the feed
material. As chicken feed, suitable particle grain sizes (d.sub.50)
are for example about 1.0 mm, about 1.2 or about 1.4 mm.
Preferably, grain size (absolute) is smaller than 10 mm. In one
embodiment, the grain size preferably is smaller than 5 mm,
particularly for grown up chicken, and even more preferably smaller
than 3 mm, as that is about the maximum size for starter laying
hens.
The feed material produced in this way is very suitable as feed
material for chicken and other fodder with a crop. Because of the
preferred low temperature during drying, extracellular enzymes like
cellulases and amylases remain active. Intake by the chicken of
such active enzymes aid in feed conversion in the crop.
The present invention also is concerned with a process for
producing feed material, comprising a) fermenting Agaricus blazei
on grain with a moisture content between 10 and 90%, until 10-50%
mycelium content, preferably on rye or oat; b) drying the fermented
grain to a moisture content of between 2 and 8 wt % (with respect
to the dry material) at a temperature below 50.degree. C., and
preferably between 20 and 45.degree. C. while applying reduced
pressure; c) and milling the so obtained fermented and dried
material to a particle size with a d.sub.50 between 0.01 and 10
mm.
The fermented, dried and optionally milled fermented grain (the
feed material) can be used as such as feed, as a separate feed
source, or it can be mixed with other common feedstock for chicken.
Preferably, the feed material is mixed with the general feedstock,
as that allows better standardization and/or reproducibility.
The feed material generally will contain between 10 and 50 wt % of
mycelium (based on dry weight), preferably between 20 and 30 wt %.
The amount of mycelium can be measured based on the content of
ergosterol. The fermentation will cause a number of useful
compounds to be present in the fermented grains. The Agaricus
blazei species produces for example, ergosterol, extra-cellular
enzymes like cellulases and amylases, and 1,3- and
1,6-.beta.-glucans.
Ergosterol is a sterol from which vitamin D is produced by the
chicken. Hence, the ergosterol--while vitamin D being important for
the calcium resorption form the gut--is thought to be instrumental
in improvement of the egg shell quality. The amount of ergosterol
in the feed material generally will be between 0.05 and 0.5 g
ergosterol/kg feed material. Ergosterol can be measured with
standard techniques, like HPLC or GC. Preferably, quantitative
extraction is performed on pulverized dry fermented grain, which
can be done with hot 80% ethanol.
It is preferred to use such an amount of feed material according
the invention, that the mixed feed comprises between 50 and 1000
microgram ergosterol per kg feed material, preferably in an amount
of 100-600 microgram ergosterol per kg feed material.
The extra-cellular enzymes, that remain active with the current
preferred drying process, may aid in an improved feed conversion.
The feed material according the present invention preferably
comprises active extra-cellular cellulases. The presence of
extra-cellular cellulases can be determined according to the
methods described in Mycobiology 39(2): 103-108 (2011). In a
preferred embodiment, the cellulase activity is between 0.1 and 0.8
unit/ml when 10 gram of material is extracted with 20 ml of water.
In this extraction, 10 gr of dry feed material is soaked with 20 ml
of water at 20.degree. C. for two hours, and thereafter the water
is squeezed out of the mixture in a press, and the enzymatic
activity in the extracted water measured.
The 1,3- and 1,6-.beta.-glucans are known to improve the immune
system. The glucans are water soluble. The amount of these glucans
generally will be between 3 to 100 gram, preferably 6 to 60 gram
and most preferable 10 to 40 gram per kg of the feed material (i.e.
the fermented grain).
The amount of 1,3- and 1,6-.beta.-glucans in the total feed for
chicken will be between 20 and 200 mg/kg feed, preferably between
50 and 100 mg/kg feed. 1,3- and 1,6-.beta.-glucans can be measured
with standard kits. However, it appeared that measurement in the
fermented grain resulted in unreliable results. Hence, the amount
of 1,3- and 1,6-.beta.-glucans in the fermented grains can be
determined indirectly, by determining these compounds in pure
fermented ABM, and calculating from the % conversion (measured via
the amount of ergosterol) the amount of 1,3- and
1,6-.mu.-glucans.
Some analysis in the fermented grain is complicated because of the
components of the grain. It can be useful to determine the
production of useful components in pure mycelium. This can be done
by growing the mycelia in a fermenter under non-limiting
conditions, removing the medium, and measuring the components of
the mycelium. Obviously, only intracellular components are measured
in this way. The ABM used by the present inventors produced about
10.7 g/kg GlcNac release, as measure for chitin, about 0.65 g/kg
mycelium of ergosterol, about 120 g/kg mycelium of
1,3-.beta.-glucans and 1,6-.beta.-glucans.
The amount of mycelium can be measured based on the content of
chitin, but also for example based on the amount of ergosterol. The
determination while using chitin is preferred, as that is a stable
compound (while ergosterol is UV sensitive). When done correctly,
both methods should give approximately the same results.
The method used for the determination of conversion via the
assessment of amount of chitin is described in Nature protocols
(2007) 1:2995-3000. In short, chitin is first liberated from the
cell walls, and while using chitinase, chitin is hydrolyzed and
N-acetylglucosamine is formed, which can be determined
quantitatively with tetraborate and Reissig reagent. The amount
found in the fermented grain should be compared with a calibration
line made from pure mycelium.
The amount of mycelium (as pure dry matter) given to the laying
hens per day, generally will be between 0.1 and 1.0 kg per ton
feed, preferably between 0.2 and 0.8 kg per ton feed. The amount of
feed material, to be added to the normal feedstock can be adjusted
after assessing the amount of mycelium. The amount of feed material
generally will be about 0.2 kg per ton or more, preferably about
0.5 kg or more, and more preferably about 1 kg per ton feed or
more. Generally, the amount will be about 10 kg per ton feed or
less, preferably about 5 kg per ton feed or less. Suitable examples
include 2, 3 or 5 kg per ton feed.
A hen consumes initially about 70 gram or more feed per day per kg
bodyweight, and after a few weeks about 100 up to 125 gram of feed
per day.
The amount of feed material (25% conversion on rye) according to
the invention given per laying hen is between about 20 and 500
milligram per kg chicken-bodyweight per day, preferably between 50
and 300 milligram per kg chicken-bodyweight per day, like for
example about 100, about 150, or about 200
milligram/chicken-bodyweight/day.
In view of the small amounts, the feed material according the
present invention preferably is mixed with other feed components.
Common feedstock for chicken are grains, soybean meal, sesame meal,
fish meal, cottonseed meal and the like. Preferably, the feedstock
comprises more than 15 wt %, like for example between 15 and 23 wt
% protein and a sufficient amount of energy. Furthermore,
preferably the feedstock comprises calcium, in an amount of about 2
wt % or more, preferably about 3 wt % or more, and more preferably
about 4 wt % or more. The feedstock preferably also comprises about
1-2 wt % linoleic acid (preferably as glycerol ester).
Additionally, the feed preferably comprises vitamins like vitamin A
(preferably more than 10000 IU), vitamin D (preferably more than
2000 IU) and a sufficient amount of biotin, cholin, antioxidants,
manganese, zinc, and trace materials. If the feed material
according to the present invention is used, the amount of vitamin D
may be lower. Generally, the feed for chicken varies over the
laying period. The feed generally contains preferably more than 15
wt % protein, more than 2 wt % calcium and more than 1 wt %
linoleic acid.
It is not necessary to use the feed material of the present
invention every day. Intermittent administration, like for example
every other day, once every three days, or once a week may be
suitable as well. It is thought that every day, or every other day
is most effective.
In one embodiment of the invention, the feed material is used
during the whole of the growth and egg laying period. Using the
feed material during the whole of the period has the advantage that
mortality in the initial phase of growth is reduced, which leads to
an increase in egg production for a given group of hens.
In another embodiment, the feed material is used only during the
second half of the laying period of the laying hens. In this
embodiment, the feed material is used only (or predominantly only;
incidental use before that period is of course immaterial) from
about week 30 onwards of the egg laying period or later, and
preferably from about week 40 onwards, and most preferably from
week 45 onwards of the egg laying period. The feed material should
however at least be used from about week 50 onwards, otherwise
reduced improvement will be observed. This embodiment has the
advantage of reduced use of feed material, while still a very
effective increase of egg laying period with good egg shell quality
is observed.
The present invention will be elucidated with the following
examples, without being limited thereto.
EXAMPLES
Feed Material
Feed material was produced as follows. Bags of 15 L with 3 kg wet
rye (50% moisture) were inoculated with about 80 mg ABM mycelium
and cultivated for 42 days at 28.degree. C. at 50% humidity. The
product was dried (to a moisture content of less than 5%) with a
vacuum rotary vapor drum at 35.degree. C. at 20 mmHg pressure. The
dried material was coarsely milled at d.sub.50 1000 micron (1
mm).
The bioconversion was indirectly measured via the ergosterol
amount. The method was as follows: The samples were lyophilized for
48 hours before extraction. Ergosterol was extracted in the dark
from 100 mg of biological material (mushroom or vegetable matrix)
in 15 ml of KOH/MeOH (1:10, w/v) for 1 h at 80.degree. C. and then
rapidly cooled in ice. 30 .mu.g of internal standard (cholesterol:
1 mg/l in hexane) are added prior extraction. After adding 2 ml of
water, an extraction is performed twice with 4 mL of hexane. The
organic layers were collected and evaporated under nitrogen flow.
Ergosterol was analyzed by gas chromatography (FID detector, column
DB-5). The amount of ergosterol in 100% ABM is for this strain 0.65
g/kg.
The ergosterol and glucan content of the feed material is reported
in Table 1; amounts are values of the feed material (the fermented
and dried grain). The amount of glucans was measured in submerged
fermented ABM, and measured in accordance with AACC method 32-23
(McClearly method), as for example described by Megazyme (Wicklow,
Ireland). The amount in the feed material was calculated based on
the conversion determined via the ergosterol content.
TABLE-US-00001 TABLE 1 Fermentation % Run (bioconversion)
Ergosterol 1,3-1,6-.beta.-glucan 1 (oat) 25% in 42 days 0.16 g/kg
30.2 g/kg 2 (oat) 18% in 35 days 0.12 g/kg 21.8 g/kg 3 (rye) 35% in
35 days 0.23 g/kg 42.4 g/kg
Trial with Laying Hens
A large scale trial was set up, with two groups of laying hens
(Lohmann Bruin Lite) of 17 week old in exactly the same buildings.
The groups consisted of 10000 and 11000 laying hens respectively.
The group of 11000 hens were treated whereas the other group was
used as a control. The management for the two groups were the same
(temperature, water, feed, air treatment etc.). For the treated
group 3 kg fermented oat (20% bioconversion; i.e. 20% by weight
mycelium) per ton feed was added by an automated dosing unit (WAM
Holland). All the feed for the treated group was enriched with the
ABM feed material. The standard feed consisted of standard
optimized feed for laying hens. This type of feed is adapted to the
laying hens over the period of egg production.
Results
Although some variations were observed during the trial with
respect to egg laying performance, the treated group overall
produced about 3% more eggs on the same amount of feed (in g/egg).
The maximum egg laying period (>93%) was about 12 weeks in the
control. In the treated group, >93% of good quality eggs was
achieved from week 31 up to a at least 60 weeks; so for more than
30 weeks.
The 75% performance parameter (where 75 out of 100 chicken lay on
the average 1 egg a day) was reached after 60 weeks egg laying
(total life span 77 weeks) for the control group. The 75% figure
for the treated group was at 67 weeks (84 weeks total life span).
So next to the 3% increase during the "normal" period, the treated
laying hens produced 350,000 eggs more in the additional 7 weeks
before slaughter (i.e. more than additional 30 eggs per hen,
leading to an average increase of about 20% in number of eggs
produced per hen). The overall mortality was about 1/4.sup.th lower
in the treated group than in the control groups.
The ABM feed material was added over the complete period i.e. 165
gram ABM feed material for the treated group in 67 weeks.
Total egg production over the 60 week egg laying period is given in
table 2:
TABLE-US-00002 TABLE 2 Birds Average number of Stable Birds start
average Eggs eggs per bird 1 (control) 10,000 birds 9,800 birds
3,498,600 357 2 (treated) 11,000 birds 10,835 birds 4,225,650
390
the average quality and weight of the eggs were essentially the
same for the two groups through a large part of the trial. The egg
shells in the treated group appear to be thicker at the end of the
laying period. The quality of the eggs in the treated group allow
for egg production up to 67 weeks, also because the egg-shell is of
sufficient quality. Further Feed Material
Further feed material was produced in a comparable manner. The
bioconversion was indirectly measured via the chitin amount as
described in Nature protocols 1, 2995-3000, 2007. The grain used
does not comprise chitin, so all chitin measured originates from
the mycelium. The conversion can be calculated via the amount of
formed GlcNac, and its absorbance at 585 nm. Per mg of dry fungal
mass, the amount of GlcNac is 1.06% (standard deviation, 0.2%).
Hence, the OD measured for an amount of GlcNac obtained from 10 mg
of pure dry mycelium is 0.93. Hence, for a measured OD (ODm),
dividing the ODm by 0.093, gives the amount of mycelium. This
amount, multiplied by 100, and divided by the initial amount of
mycelium for a given sample gives the percentage bioconversion. The
feed material, produced on rye showed a bioconversion of 26% in 70
days. The amount of ergosterol was 0.17 g/kg and the amount of
.beta.-glucans was 31 g/kg.
Trial with Laying Hens During Part of the Egg Laying Period
In a further trial, feed material was used only from week 41
onwards, hence during the last 20 weeks of the egg laying period of
untreated hens. The amount used was twice the amount given in the
former trial (hence, 5 kg of the product with 26% bioconversion)
mixed with the normal feed for laying hens as described in the
example above. The egg laying period was extended with 5 weeks till
the threshold value of 75% was reached, with eggs with sufficient
egg shell quality.
* * * * *
References